Objective To study the application of multi‐tumor marker protein chip for early tumor screening and diagnosis . Methods From Nov .2011 to Dec .2015 ,10 736 samples ,including people receiving physical examination and with high risk of canc‐er in Fengcheng Hospital were collected .Twelve tumor markers in serum(AFP ,CEA ,NSE ,CA125 ,CA153 ,CA242 ,CA199 ,PSA ,f‐PSA ,FER ,β‐HCG and HGH)were measured by multi‐tumor markers protein chip detective system ,and the results were analyzed . Results We found out 967 samples with positive markers in the 10 736 samples .Of which ,496 were male and 471 were female ,the positive rate were 4 .62% and 4 .39% respectively .Totally 473 were diagnosed with tumor confirming by clinical pathology ,postive diagnosis rate was 48 .91% .Conclusion The multi‐tumor marker protein chip (C12 system) can detect multiple tumor markers simultaneously to improve screening process and achieve rapid detection ,w hich has higher positive detectiong rate and clinical value on diagnosing malignant tumor in early stage .

Objective To explore the preoperative assessment of airway morphology affecting the early-phase prognosis by observing congenital tracheal stenosis survival after surgical treatment.Methods Totally 52 cases of CTS surgically treated in the Shanghai children's Medical Center,from April 2007 to June 2013,were retrospectively analyzed to study their clinical characteristics,survival condition and related factors influencing the early-phase prognosis.Using log-rank test and Cox multiple factors analysis for statistical analysis.Results Overall postoperative survival rate of 3 months was 73.1％.There were 14 deaths among 48 patients.Single factor and multiple factors analysis showed that the cross-sectional area(CSA) of stenosis and Anton-Pacheco classification were both associated with early-phase prognosis as independent factors(P ＜0.05).Long CPB duration was a prognostic factor for the outcome of surgical treatment for CTS(P ＜ 0.05).CPB time was significantly longer in death cases than in surviving cases.Conclusion CSA and Anton-Pacheco classification were significant prognostic factors for CTS.The proper utilization of radiologic imaging allow for improved patient care.

Objective To determine the population structure of Streptococcus pncumoniae isolates collected from infants with eye infections,including drug resistance,resistance genes, serotypes and molecular types.Methods The susceptibility of 39 isolates to 10 antibacterial agents was tested by K-B disk diffusion and Etest.Latex agglutination test was performed to determine the serotype of the strains,and PCR was carried out to detect macrolides resistance genes mefE and ermB.Molecular types of the 20 strains were determined by multilocus sequence typing (MLST).Results A total of 39 Streptococcus pneumoniae isolates were obtained,in which 30 (76.9％) were resistant to 3 or more antibacterial agents,and no vancomycin,penicillin or cefotaxime resistant strain was found.ermB gene was found in 33 strains and mefE gene was found in 4 strains.Twelve serotypes were found,and the most frequent serotypes were 19 (8/39) and 14 (4/39). Seventeen strains (43.6％) were covered in PCV7 vaccine. The international clone Taiwan19F-14 and Spain23F-1 were found by MLST. Conclusions Streptococcus pneumoniae isolates from infants with eye infections include international resistance clones.The distribution of serotype and molecular type are dispersed, and the clones are sporadic. The isolates are highly resistant to commonly used antibacterial agents.

Objective To present a miniaturized nucleic acid amplification system for spot rapid detection .Methods A miniaturized nucleic acid amplification system with structured packed porous media of particles to uniform the air temperature was designed according to the working principle and heat transfer characteristics of an air -heated nucleic acid amplification system.Thermodynamic simulation and temperature cycling test were carried on to verify the feasibility of the system.Results The structured packed porous media of particles worked well in uniforming the air temperature of the system and the temperature uniformity could reach 0.8℃.The miniaturized nucleic acid amplification system with a volume parameter of 80 mm ×40 mm ×20 mm(length ×height ×width)was portable.The average rate of heating was 10℃/s while the average rate of cooling was 5℃/s.Compared with standard PCR instrument , the miniaturized nucleic acid amplification system performed well in the process of amplification and met the requirements of preliminary design .Conclusion The miniaturized nucleic acid amplification system with a rapid reaction velocity and portable volume could be applied to nucleic acid detection of unknown samples on the spot .

Objectives To study the performance of different microbial automated inoculation systems and to evaluate the performance of the Probact microbial automated inoculation and incubation system ( Probact system) and its applications in clinical microbiology laboratory.Methods A total of 160 clinical specimens, including respiratory secretions ( n=61 ) , urine ( n=49 ) , and feces ( n=50 ) , that were submitted to the Clinical Microbiology Laboratory in Peking Union Medical College Hospital of Chinese Academy of Medical Sciences from February 2015 to April 2015 were evaluated.These specimens were processed with conventional manual method, the Probact automated inoculation system, and PREVI Isola Inoculator.The quantity of bacterial species recovery, number of effectively isolated colonies, total number of colonies recovery per plate, and time of processing the 160 specimens by the three methods were evaluated. Wilcoxon signed-rank test and Kruskal-Wallis rank sum test were used for statistical analysis.Results The Probact system had significantly higher quantity of bacterial species recovery (respiratory specimens 3.41 ±1.40, urine 1.92 ±0.86, and feces 1.16 ±0.79) than those by the Isola Inoculator (respiratory specimens 3.75 ±1.29, urine 2.24 ±0.97, and feces 1.92 ±0.72), (P=0.006, 0.011, <0.001).Compared to the manual method, Probact performed less quantity of bacterial species recovery for respiratory specimens(3.85 ±1.38), but higher in feces(0.80 ±0.81)( P<0.001).There is no significant differences for urine ( 1.84 ±1.23 ) ( P=0.266 ) .As for number of isolated colony, the Probact system ( respiratory specimens 12.16 ±7.72, urine 2.71 ±4.24, and feces 5.40 ±5.04 ) had significant smaller numbers than that of Isola Inoculator (respiratory specimens 16.56 ±5.76, urine 4.35 ± 4.89, and feces 8.40 ±3.70) (P<0.001,0.007,0.003).However, both system had larger numbers of isolated colonies than those by the manual method (respiratory specimens 11.30 ±8.42, urine 2.67 ±4.34, and feces 1.90 ±3.90) and the difference was significant for fecal specimens(P<0.001).Regarding the total number of colonies recovery, larger number was found by Isola Inoculator than that by the Probact system for fecal specimens, however, there were no significant differences for respiratory or urine specimens (P=0.524,0.738).Compared with manual method, the Probact system had significantly more numbers of colonies recovery for respiratory and fecal specimens ( P<0.001 ) . The total time for processing 160 specimens was shortest for manual method (281 min), followed by Probact system (419 min) and Isola Inoculator (495 min) .Conclusions The performance of the Probact system is better than the manual method but no superior to the Isola Inoculator.The Probact system can meet the clinical need in terms of full automation and standardization of specimen inoculation and prevention of bias of processing by laboratory staffs using manual method.

Objective: To explore the application value on combined examination of blood levels of growth differentiation factor-15 (GDF-15) and NT-pro B-type natriuretic peptide (NT-proBNP) in patients after successful cardiopulmonary resuscitation (CPR) for their recent prognosis.
Methods: A total of 102 patients with sudden cardiac arrest and successful CPR in our hospital were enrolled. Blood levels of GDF-15 were examined at immediately, 12 h and 24-48 h after CPR respectively. According to GDF-15 levels, the patients were divided into 3 groups: Group A, the patients with GDF-15<1200 ng/L at all-time points,n=31; Group B, GDF-15 level consistently increasing and GDF-15>1200 ng/L at all-time points,n=35; Group C, GDF-15 level consistently increasing at 12 h and 24-48 h after CPR, while it was lower at 24-48 h than 12 h after CPR,n=36. Blood levels of NT-proBNP and left ventricular ejection fraction (LVEF) were also examined. The patients were followed-up for 6 months for post-CPR death.
Results: Blood levels of GDF-15 and NT-proBNP were related, NT-proBNP level was changing with GDF-15 varying. GDF-15 and NT-proBNP level was negatively related to LVEF (r=-0.530,P<0.001), the patients with GDF-15>1800 ng/L and NT-proBNP>400 pg/ml had the higher mortality than those had the lower levels of GDF-15 and NT-proBNP,P<0.05. Survival analysis presented that 6 months survival rate in Group B was lower than Group A and Group C,P<0.05; survival rate was similar between Group A and Group C,P>0.05.
Conclusion: Combined examination for blood levels of GDF-15 and NT-proBNP may better predict the recent prognosis in patients who received CPR.

Objective:To explore the correlation of apparent diffusion coefficient (ADC) of magnetic resonance diffusion weighted imaging (DWI) examination before radiotherapy in patients with advanced cervical squamous cell carcinoma with clinicopathological characteristics and radiotherapy efficacy.Methods:The clinical data of 80 patients with advanced cervical cancer who were admitted to the Second Hospital of Nanjing from September 2019 to March 2022 were retrospectively analyzed. All patients underwent magnetic resonance imaging (MRI) DWI examination. The differences in ADC values among cervical squamous cell carcinoma patients with different clinicopathological characteristics were analyzed. The patients were divided into the effective group (complete remission+partial remission) and the ineffective group (stable disease+progressive disease) based on the radiotherapy effect, and the differences in ADC values between the two groups were compared. The logistic regression model was used to analyze the factors affecting the radiotherapy efficacy of patients with advanced cervical squamous cell carcinoma.Results:Among 80 patients with advanced cervical squamous cell carcinoma, 21 achieved complete remission, 31 achieved partial remission, 25 achieved stable disease, and 3 achieved progressive disease after radiotherapy; there were 52 cases in the effective group and 28 cases in the ineffective group. The ADC value of the effective group before radiotherapy was higher than that of the ineffective group [(0.99±0.14)×10 -3mm 2/s vs. (0.76±0.20)×10 -3mm 2/s], and the difference was statistically significant ( t = 6.01, P < 0.001); after radiotherapy, the ADC value of the effective group was also higher than that of the ineffective group [(1.43±0.25)×10 -3mm 2/s vs. (1.11±0.23)×10 -3mm 2/s), and the difference was statistically significant ( t = 5.61, P < 0.001); the ADC values of both the effective and ineffective groups increased after radiotherapy compared to before radiotherapy (both P < 0.05). The ADC values of patients with different International Federation of Obstetrics and Gynecology (FIGO) stage, degree of pathological differentiation, depth of lesion infiltration, Ki-67 expression, lymph node metastasis, and distant metastasis were statistically significant (all P < 0.05). The results of multivariate logistic regression analysis showed that ≥FIGO stage Ⅲ, low differentiation, lymph node metastasis, lymphatic vessel infiltration, distant metastasis, and low ADC value before radiotherapy were independent risk factors for efficacy of radiotherapy in patients with advanced cervical squamous cell carcinoma (all P < 0.05). Conclusions:The ADC value before radiotherapy is a factor that affects the radiotherapy effect of patients with advanced cervical squamous cell carcinoma. The lower the ADC value before radiotherapy is, the worse the radiotherapy effect of patients will be.

Objective To explore the human papilomavirus(HPV)genotypes and epithelial thickness of invisible cervical intraepithelial neoplasia Ⅲ(CIN Ⅲ)under colposcopy. Methods One hundred and sixty-nine biopsies from 93 patients with a final diagnosis of CIN Ⅲwere extracted from the Shenzhen cervical cancer screening trialⅡ(SHENCCASTⅡ).The SHENCCASTⅡwas conducted from 2009 to 2010.All the cervical blocks from these patients were re-cut and placed on 6 slides,i.e.sandwich model, with the top and bottom sections being stained with HE, the top second be processed for other studies, 3 sections for HPV genotypes by matrix-assisted laser desorption ionization-time of flight-mass spectrometry(MALDI-TOF-MS)assay.The thickness of squamous epithelium of CINⅢwas measured by a microscope(×10)after re-cut. Colposcope directed CIN Ⅲ biopsies positively was defined as visible CIN Ⅲ, while random CIN Ⅲ biopsies positively was defined as invisible CIN Ⅲ. Results HPV16 positivity was 37.2%(16/43)and 55.6%(70/126)between invisible and visible CIN Ⅲ biopsies, respectively(χ2=4.318,P=0.038).Forty-nine cases of the 93 CINⅢpatients were HPV16 positive,while 44 of them non-HPV16 positive. The proportion of patients with ≥45 years of age for other non-HPV16 positive 40.9%(18/44)was significantly higher than that HPV16 positive 20.4%(10/49; χ2=4.630, P=0.031).Patients with HPV16 positive were more likely to have lesions ≥1 quadrant(χ2=7.786,P=0.005) than other non-HPV16 positive. Compared the average epithelium thickness of invisible CIN Ⅲ tissue (140±12)μm,the average epithelium thickness of visible CIN Ⅲtissue(161±9)μm was thicker.There was statistical difference between two groups(t=4.383,P=0.038).The mean average epithelial thickness of CIN Ⅲwith HPV16 positive(172±11)μm was thicker than that the mean average epithelial thickness of CIN Ⅲ with non-HPV16 positive(130±10)μm(t=4.784,P=0.031). Conclusions Invisible lesions is difficult to identify under colposcopy and is related to non-HPV16 positive, small lesion size and thinner squamous epithelium. For non-HPV16 positive or older women should be performed colposcope directed biopsies and randomly multi-sites biopsies by colopscopy,which may be helpful to improve the detection of CINⅢand to reduce miss diagnosis.

Objective:To investigate the 10-year outcome and prognostic factors of laparo-scopic D 2 radical distal gastrectomy for locally advanced gastric cancer. Methods:The retrospec-tive cohort study was conducted. The clinicopathological data of 652 patients with locally advanced gastric cancer who were admitted to 16 hospitals from the multicenter database of laparoscopic gastric cancer surgery in the Chinese Laparoscopic Gastrointestinal Surgery Study (CLASS) Group, including 214 cases in the First Affiliated Hospital of Army Medical University, 191 cases in Fujian Medical University Union Hospital, 52 cases in Nanfang Hospital of Southern Medical University, 49 cases in West China Hospital of Sichuan University, 43 cases in Xijing Hospital of Air Force Medical University, 25 cases in Jiangsu Province Hospital of Chinese Medicine, 14 cases in the First Medical Center of the Chinese PLA General Hospital, 12 cases in No.989 Hospital of PLA, 12 cases in the Third Affiliated Hospital of Sun Yat-Sen University, 10 cases in the First Affiliated Hospital of Nanchang University, 9 cases in the First People's Hospital of Foshan, 7 cases in Zhujiang Hospital of Southern Medical University, 7 cases in Fujian Medical University Cancer Hospital, 3 cases in Zhongshan Hospital of Fudan University, 2 cases in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 2 cases in Peking University Cancer Hospital & Institute, from February 2004 to December 2010 were collected. There were 442 males and 210 females, aged (57±12)years. All patients underwent laparoscopic D 2 radical distal gastrectomy. Observation indicators: (1) surgical situations; (2) postoperative pathological examination; (3) postoperative recovery and complications; (4) follow-up; (5) prognostic factors analysis. Follow-up was conducted by outpatient examination and telephone interview to detect the tumor recurrence and metastasis, postoperative survival of patients up to March 2020. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M( Q1, Q3) or M(range). Count data were described as absolute numbers or percen-tages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data was analyzed using the rank sum test. The life table method was used to calculate survival rates and the Kaplan-Meier method was used to draw survival curves. Log-Rank test was used for survival analysis. Univariate and multivariate analyses were analyzed using the COX hazard regression model. Results:(1) Surgical situations: among 652 patients, 617 cases underwent D 2 lymph node dissection and 35 cases underwent D 2+ lymph node dissection. There were 348 cases with Billroth Ⅱ anastomosis, 218 cases with Billroth Ⅰ anastomosis, 25 cases with Roux-en-Y anastomosis and 61 cases with other digestive tract reconstruction methods. Twelve patients had combined visceral resection. There were 569 patients with intraoperative blood transfusion and 83 cases without blood transfusion. The operation time of 652 patients was 187(155,240)minutes and volume of intraoperative blood loss was 100(50,150)mL. (2) Postoperative pathological examina-tion: the maximum diameter of tumor was (4.5±2.0)cm of 652 patients. The number of lymph node dissected of 652 patients was 26(19,35), in which the number of lymph node dissected was >15 of 570 cases and ≤15 of 82 cases. The number of metastatic lymph node was 4(1,9). The proximal tumor margin was (4.8±1.6)cm and the distal tumor margin was (4.5±1.5)cm. Among 652 patients, 255 cases were classified as Borrmann type Ⅰ-Ⅱ, 334 cases were classified as Borrmann type Ⅲ-Ⅳ, and 63 cases had missing Borrmann classification data. The degree of tumor differentiation was high or medium in 171 cases, low or undifferentiated in 430 cases, and the tumor differentiation data was missing in 51 cases. There were 123, 253 and 276 cases in pathological stage T2, T3 and T4a, respectively. There were 116, 131, 214 and 191 cases in pathological stage N0, N1, N2 and N3, respectively. There were 260 and 392 cases in pathological TNM stage Ⅱ and Ⅲ, respectively. (3) Postoperative recovery and complications: the time to postoperative first out-of-bed activities, time to postoperative first flatus, time to the initial liquid food intake, duration of postoperative hospital stay of 652 patients were 3(2,4)days, 4(3,5)days, 5(4,6)days, 10(9,13)days, respectively. Among 652 patients, 69 cases had postoperative complications. Clavien-Dindo grade Ⅰ-Ⅱ, grade Ⅲa, grade Ⅲb, and grade Ⅳa complications occurred in 60, 3, 5 and 1 cases, respectively (some patients could have multiple complications). The duodenal stump leakage was the most common surgical complication, with the incidence of 3.07%(20/652). Respiratory complication was the most common systemic complication, with the incidence of 2.91%(19/652). All the 69 patients were recovered and discharged successfully after treatment. (4) Follow-up: 652 patients were followed up for 110-193 months, with a median follow-up time of 124 months. There were 298 cases with postoperative recurrence and metastasis. Of the 255 patients with the time to postoperative recurrence and metastasis ≤5 years, there were 21 cases with distant metastasis, 69 cases with peritoneal metastasis, 37 cases with local recurrence, 52 cases with multiple recurrence and metastasis, 76 cases with recurrence and metastasis at other locations. The above indicators were 5, 9, 10, 4, 15 of the 43 patients with the time to postoperative recurrence and metastasis >5 years. There was no significant difference in the type of recurrence and metastasis between them ( χ2=5.52, P>0.05). Cases in pathological TNM stage Ⅱ and Ⅲ were 62 and 193 of the patients with the time to postoperative recurrence and metastasis ≤5 years, versus 23 and 20 of the patients with the time to postoperative recurrence and metastasis >5 years, showing a significant difference in pathological TNM staging between them ( χ2=15.36, P<0.05). Cases in pathological stage T2, T3, T4a were 42, 95, 118 of the patients with the time to postoperative recurrence and metastasis ≤5 years, versus 9, 21, 13 of the patients with the time to postoperative recurrence and metastasis >5 years, showing no significant difference in pathological T staging between them ( Z=-1.80, P>0.05). Further analysis showed no significant difference in cases in pathological stage T2 or T3 ( χ2=0.52, 2.08, P>0.05) but a significant difference in cases in pathological stage T4a between them ( χ2=3.84, P<0.05). Cases in pathological stage N0, N1, N2, N3 were 19, 44, 85, 107 of the patients with the time to postoperative recurrence and metastasis ≤5 years, versus 12, 5, 18, 8 of the patients with the time to postoperative recurrence and metastasis >5 years, showing a significant difference in pathological N staging between them ( Z=-3.34, P<0.05). Further analysis showed significant differences in cases in pathological stage N0 and N3 ( χ2=16.52, 8.47, P<0.05) but no significant difference in cases in pathological stage N1 or N2 ( χ2=0.85, 1.18, P>0.05). The median overall survival time was 81 months after surgery and 10-year overall survival rate was 46.1% of 652 patients. The 10-year overall survival rates of patients in TNM stage Ⅱ and Ⅲ were 59.6% and 37.5%, respectively, showing a significant difference between them ( χ2=35.29, P<0.05). In further analysis, the 10-year overall survival rates of patients in pathological TNM stage ⅡA, ⅡB, ⅢA, ⅢB and ⅢC were 65.6%, 55.8%, 46.9%, 37.1% and 24.0%, respectively, showing a significant difference between them ( χ2=55.06, P<0.05). The 10-year overall survival rates of patients in patholo-gical stage T2, T3 and T4a were 55.2%, 46.5% and 41.5%, respectively, showing a significant difference between them ( χ2=8.39, P<0.05). The 10-year overall survival rates of patients in patholo-gical stage N0, N1, N2 and N3 were 63.7%, 56.2%, 48.5% and 26.4%, respectively, showing a signifi-cant difference between them ( χ2=54.89, P<0.05). (5) Prognostic factors analysis: results of univariate analysis showed that age, maximum diameter of tumor, degree of tumor differentiation as low or undifferentiated, pathological TNM staging, pathological T staging, pathological stage N2 or N3, post-operative chemotherapy were related factors for the 10-year overall survival rate of locally advanced gastric cancer patients undergoing laparoscopic D 2 radical distal gastrectomy ( hazard ratio=1.45, 1.64, 1.37, 2.05, 1.30, 1.68, 3.08, 0.56, 95% confidence interval as 1.15-1.84, 1.32-2.03, 1.05-1.77, 1.62-2.59, 1.05-1.61, 1.17-2.42, 2.15-4.41, 0.44-0.70, P<0.05). Results of multivariate analysis showed that maximum diameter of tumor >4 cm, low-differentiated or undifferentiated tumor, pathological TNM stage Ⅲ were independent risk factors for the 10-year overall survival rate of locally advanced gastric cancer patients undergoing laparoscopic D 2 radical distal gastrectomy ( hazard ratio=1.48,1.44, 1.81, 95% confidence interval as 1.19-1.84, 1.11-1.88, 1.42-2.30, P<0.05) and postoperative chemotherapy was a independent protective factor for the 10-year overall survi-val rate of locally advanced gastric cancer patients undergoing laparoscopic D 2 radical distal gastrec-tomy ( hazard ratio=0.57, 95% confidence interval as 045-0.73, P<0.05). Conclusions:Laparoscopic assisted D 2 radical distal gastrectomy for locally advanced gastric cancer has satisfactory 10-year oncologic outcomes. A high proportion of patients in pathological TNM stage Ⅲ, pathological stage T4a, pathological stage N3 have the time to postoperative recurrence and metastasis ≤5 years, whereas a high proportion of patients in pathological TNM stage Ⅱ or pathological stage N0 have the time to postoperative recurrence and metastasis >5 years. Maximum diameter of tumor >4 cm, low-differentiated or undifferentiated tumor, pathological TNM stage Ⅲ are independent risk factors for the 10-year overall survival rate of locally advanced gastric cancer patients undergoing laparos-copic D 2 radical distal gastrectomy. Postoperative chemotherapy is a independent protective factor for the 10-year overall survival rate of locally advanced gastric cancer patients undergoing laparos-copic D 2 radical distal gastrectomy.

The CRISPR/Cas9 based prime editing (PE) technique enables all 12 types of base substitutions and precise small DNA deletions or insertions without generating DNA double-strand breaks. Prime editing has been successfully applied in plants and plays important roles in plant precision breeding. Although plant prime editing (PPE) can substantially expand the scope and capabilities of precise genome editing in plants, its editing efficiency still needs to be further improved. Here, we review the development of PPE technique, and introduce structural composition, advantages and limitations of PPE. Strategies to improve the PPE editing efficiency, including the T_m-directed PBS length design, the RT template length, the dual-pegRNA strategy, the PlantPegDesigner website, and the strategies for optimizing the target proteins of PPE, were highlighted. Finally, the prospects of future development and application of PPE were discussed.
CRISPR-Cas Systems/genetics* ; DNA ; Gene Editing ; Genome, Plant/genetics* ; Plant Breeding ; Plants/genetics*